LOCUS       sequence-435135-        7911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7911)
  AUTHORS   Stutz A, Horvath GL, Monks BG, Latz E
  TITLE     ASC speck formation as a readout for inflammasome activation.
  JOURNAL   Methods Mol Biol. 2013;1040:91-101. doi: 
            10.1007/978-1-62703-523-1_8.
  PUBMED    23852599
REFERENCE   2  (bases 1 to 7911)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods Mol
            Biol."; date: "2013"; volume: "1040:91-101. doi"; pages: " 
            10.1007/978-1-62703-523-1_8"
COMMENT     Sequence Label: pRP_ASC-LmCerulean
FEATURES             Location/Qualifiers
     source          1..7911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        215..594
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        595..798
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             799..974
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    1037..1236
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1437..1853
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1783..1805
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1821..1837
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1860..2459
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1860..1879)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2356..2376
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     enhancer        2599..2978
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2979..3182
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3179..3203
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(3944..3946,3947..3949,3950..4663)
                     /codon_start=1
                     /product="monomeric super enhanced variant of CFP (Matsuda 
                     et al., 2008)"
                     /label=mseCFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNDISDNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3989..4010)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4250..4269)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4597..4618
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4717..5310
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5393..5413)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5411..5708
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5578..5713
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5640..5659
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(5754..5770)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5754..5770)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5767..5789)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5778..5794
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5802..5808,5809..5826,5827..5832))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5847..5868
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5985..6002)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6156..6744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6236..6255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6915..7706,7707..7775))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7538..7557
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7776..7880)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg agttgacatt gattattgac tagttattaa
      241 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      301 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      361 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      421 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      481 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      541 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      601 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      661 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      721 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      781 gtctatataa gcagagctca ataaaagagc ccacaacccc tcactcggcg cgccagtctt
      841 ccgatagact gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc tgtttgcatc
      901 cgaatcgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccacgac
      961 gggggtcttt catttggggg ctcgtccggg atttggagac ccctgcccag ggaccaccga
     1021 cccaccaccg ggaggtaagc tggccagcaa cttatctgtg tctgtccgat tgtctagtgt
     1081 ctatgtttga tgttatgcgc ctgcgtctgt actagttagc taactagctc tgtatctggc
     1141 ggacccgtgg tggaactgac gagttctgaa cacccggccg caaccctggg agacgtccca
     1201 gggactttgg gggccgtttt tgtggcccga cctgaggaag ggagtcgatg tggaatccga
     1261 ccccgtcagg atatgtggtt ctggtaggag acgagaacct aaaacagttc ccgcctccgt
     1321 ctgaattttt gctttcggtt tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca
     1381 gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aattagggcc
     1441 agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
     1501 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
     1561 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
     1621 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca
     1681 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
     1741 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
     1801 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gcctccggta
     1861 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     1921 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     1981 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     2041 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     2101 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     2161 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     2221 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     2281 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     2341 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     2401 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag
     2461 gatcgatccg gccattagcc atattattca ttggttatat agcataaatc aatattggct
     2521 attggccatt gcatacgttg tatccatatc ataatatgta catttatatt ggctcatgtc
     2581 caacattacc gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     2641 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     2701 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     2761 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     2821 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     2881 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     2941 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     3001 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     3061 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     3121 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     3181 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     3241 gaagacaccg ggaccgatcc agcctccgcg gccccaagct tccgcgtacg cccgaattcc
     3301 aggatctatg gggcgcgcgc gcgacgccat cctggatgcg ctggagaacc tgaccgccga
     3361 ggagctcaag aagttcaagc tgaagctgct gtcggtgccg ctgcgcgagg gctacgggcg
     3421 catcccgcgg ggcgcgctgc tgtccatgga cgccttggac ctcaccgaca agctggtcag
     3481 cttctacctg gagacctacg gcgccgagct caccgctaac gtgctgcgcg acatgggcct
     3541 gcaggagatg gccgggcagc tgcaggcggc cacgcaccag ggctctggag ccgcgccagc
     3601 tgggatccag gcccctcctc agtcggcagc caagccaggc ctgcacttta tagaccagca
     3661 ccgggctgcg cttatcgcga gggtcacaaa cgttgagtgg ctgctggatg ctctgtacgg
     3721 gaaggtcctg acggatgagc agtaccaggc agtgcgggcc gagcccacca acccaagcaa
     3781 gatgcggaag ctcttcagtt tcacaccagc ctggaactgg acctgcaagg acttgctcct
     3841 ccaggcccta agggagtccc agtcctacct ggtggaggac ctggagcgga gcctcgagag
     3901 tacagtacca agggctcgtg accctccagt cgctaccatc gagatggtga gcaagggcga
     3961 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     4021 caggttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     4081 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     4141 ctggggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     4201 gtccgccatg cccgaaggct acgtccagga gcgtaccatc ttcttcaagg acgacggcaa
     4261 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     4321 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacga
     4381 catcagcgac aacgtctata tcaccgccga caagcagaag aacggcatca aggcccactt
     4441 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     4501 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     4561 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     4621 cgccgccggg atcactctcg gcatggacga gctgtacaag taagcggccg catcgataaa
     4681 ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt
     4741 tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat
     4801 agagaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc caaacaggat
     4861 atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa cagctgaata
     4921 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     4981 tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga tgtttccagg
     5041 gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc
     5101 tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac
     5161 tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc caataaaccc
     5221 tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga
     5281 ttgactaccc gtcagcgggg gtctttcatt tgggggctcg tccgggatcg ggagacccct
     5341 gcccagggac caccgaccca ccaccgggag gtaagctggg aactaattct gtggaatgtg
     5401 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5461 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     5521 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     5581 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5641 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     5701 ttttttggag gcctagtacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     5761 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5821 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5881 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5941 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     6001 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     6061 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     6121 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6181 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6241 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6301 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6361 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6421 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6481 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6541 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     6601 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6661 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6721 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6781 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6841 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6901 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6961 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7021 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7081 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7141 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7201 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7261 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7321 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7381 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7441 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7501 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7561 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7621 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7681 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7741 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7801 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7861 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
